Enzyme-catalyzed peptide synthesis in biphasic aqueous-organic systems

N-Terminal Protein Modification Using Simple Aminoacyl Transferase Substrates

“…16,28–37 Reverse proteolysis methods can be used to modify the N-terminus under conditions that do not require protein unfolding, but the reaction can be difficult to drive to completion without high protein concentrations. 38–51 Several other chemoenzymatic methods are valuable in functioning under benign conditions, but have moderate-sized target sequences which must be appended to the protein. 52–59 Here, we describe a minimal system for N-terminal protein labeling that utilizes only adenonsine esters of natural or unnatural aminoacids and a single, readily available enzyme.…”

Miniaturized biocatalysis on polyacrylate‐based capillary monoliths

“…Although it has already been bound to various supports, the use of porous polymeric monoliths that allow for a high rate of substrate perfusion between the fixed enzyme and the substrate has been investigated less 6, 9–11, 33–36. However, the immobilization of proteases for peptide synthesis purposes has been investigated, for example, for carboxypeptidase Y and chymotrypsin 37–39. In addition to the hydrolysis of proteins and peptides, serine proteases can catalyze acyl‐transfer reactions from acyl donors to various acceptors 40.…”

Successful Application of the Lipase as a Catalyst in the Combinative Approach, Using an Efficient and Selective Iterative Procedure for the Chemo-Enzymatic Alpha-Amino Group Protection in the Natural Amino Acids, Representing the Ornithyl Model System as a Starting Research Pattern: Homogeneous and Heterogeneous Synthesis

“…To achieve a lowcost industrial preparation of chiral active pharmaceutical products, the enzyme should be used in many times without its complicated regeneration and sharply lose of its native structure and activity. The alternative using of immobilized enzyme in water-organic media is complicated, due to the difficult and prolonged separation of the immobilized enzyme from the another components in the reaction mixture [20], as well as because of the slow reagents diffusion, and finally -due to the low and poor enzyme activity -many times longer reaction time is required [20,21]. Moreover, some methodologies for enzyme immobilization require expensive matrices utilization and a specific methodology for immobilization: 1) adsorption to the outside of an inert material -on glass, alginate beads or matrix; 2) entrapment in insoluble beads or microspheres, such as calcium alginate beads; 3) covalently cross-linkage of the enzyme to a matrix through a chemical interaction by a spacer molecule like poly(ethylene glycol), that helps reduce the steric hindrance by the substrate.…”